Pharmaceutical composition including 1,2-naphthoquinone derivative compound for prevention or treatment of solid cancers or blood cancers

ABSTRACT

The present invention relates to a pharmaceutical composition for the prevention or treatment of solid cancers or blood cancers such as acute leukemia or chronic leukemia, including, as an active ingredient, a 1,2-naphthoquinone derivative compound or a pharmaceutically acceptable salt thereof, wherein the 1,2-naphthoquinone derivative compound has excellent effects in killing cancer cells of various solid cancers, acute leukemia, and chronic leukemia, and thus, can be useful as a pharmaceutical composition for the prevention or treatment of cancer, in particular, solid cancers, acute leukemia, or chronic leukemia.

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application claims the benefit of Korean Patent Application Nos.10-2018-0040913, 10-2018-0040884 and 10-2018-0040895 filed on Apr. 9,2018 with the Korean Intellectual Property Office, the disclosures ofwhich are incorporated herein by reference in their entirety.

TECHNICAL FIELD

The present invention relates to a pharmaceutical composition for theprevention or treatment of solid cancers or blood cancers such as acuteleukemia or chronic leukemia, the pharmaceutical composition including a1,2-naphthoquinone derivative compound.

BACKGROUND ART

Cells, which are the smallest units constituting the body, divide andgrow by the regulatory function of the cells themselves under normalconditions, and when the lifespan is exhausted or the cells are damaged,they die by themselves and maintain the balance of whole numbers.However, when problems occur in the regulatory function of the cellsthemselves due to various causes, abnormal cells that must die normallybecome overgrown, and in some cases, it invades surrounding tissues andorgans to form a mass (lump) and destroys or deforms an existingstructure, and this condition can be defined as cancer.

Cancer is one of the intractable diseases that human beings have tosolve, and huge capital has been invested in the development to treatcancer worldwide, and medical technology is also developinginnovatively. Nevertheless, deaths from cancer tends to persistentlyincrease. According to the announcement of the National StatisticalOffice, in the case of cancer patients in Korea, it was reported thatabout 220,000 new cancer patients have occurred on the basis of 2012.This number has increased about twice as much as the number of newcancer patients that occurred in 2002, and it can be seen that thenumber of cancer patients is increasing rapidly every year. However,among about 220,000 cancer patients, about 70,000 are dying from cancer,so the development of therapeutic agents for treatment of cancer isurgent.

Currently, therapies for cancer patients relies on surgery, radiationtherapy, and chemotherapy (administrating about 40 kinds of cytotoxicanticancer substances showing strong cytotoxity), but most of thesetherapies are also limited to early-stage cancer patients or specificcancers, so deaths from cancer are still increasing.

On the other hand, leukemia is divided into acute and chronic dependingon the degree of differentiation of cells, that is, the rate ofdeterioration, and is divided into myelogenous and lymphocytic dependingon the origin of the cell. Thus, it is classified into acute myeloidleukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, andchronic lymphocytic leukemia.

Acute leukemia is a disease in which abnormal leukocyte progenitor cellsor platelet progenitor cells are excessively formed due to bone marrowabnormalities. When myeloid cells proliferate, it is called acutemyeloid leukemia (AML), and when lymphoid cells proliferate, it iscalled acute lymphocytic leukemia (ALL). Because abnormal white bloodcells increase and occupy the place where hematopoiesis occurs, normalwhite blood cells, red blood cells, platelets and the like are notformed, so infection or bleeding easily occurs. If left untreated, theydie within a few months. Recently, due to the development ofchemotherapy, the viability of acute leukemia in infants hassignificantly improved, but the viability is still low in adults.

Acute myeloid leukemia is a malignant tumor arising from stem cells ofnon-lymphoid or myeloid leukocytes made in non-lymphocytic or bonemarrow, and is a hematopoietic tumor in which a gene mutation occurs ina hematopoietic mother cell, myeloid progenitor cells stopdifferentiating at various stages and thus, immature myeloblastsproliferate into a monoclonal group. It shows bone marrow dysfunctionsymptoms such as anemia, fever, increased infectivity, and bleedingtendency, and may also show symptoms of organ infiltration with tumorcells such as spleen enlargement and lymphadenopathy.

Acute lymphocytic leukemia is a blood cancer that develops in thelymphocyte lineage cells in the blood and bone marrow, and is known tobe caused by mutations in several genes involved in the process ofproliferation, differentiation, maturation and destruction of lymphocytelineage cells. The cause of the genetic mutation is currently notclearly identified, but it is estimated that genetic predisposition,viruses, multiple carcinogens, and ionizing radiation are involved as inother cancers. Symptoms observed in acute lymphocytic leukemia aresymptoms occurring while abnormal leukemia cells impair the process bywhich normal blood cells are formed, or while abnormal leukemia cellsinvade organs such as the lymph nodes, spleen, liver, brain, and spinalcord, similarly to other leukemias.

On the other hand, chronic myelogenous leukemia is caused by aphiladelphia chromosome, resulting from a transition phenomenon in whicha certain portion of human chromosome 9 and chromosome 22 is cut andthen the two cuttings change and move their positions with each other.It is a disease caused by excessive proliferation of abnormal cells inthe bone marrow while clones of the hematopoietic stem cell havingphiladelphia chromosome expand abnormally. Because this disease accountsfor about 25% of all adult leukemias and occurs frequently in the 30s to50s elderly, it is also called adult leukemia, but it can occur in allage groups and also occurs even in children or adolescents.

The Philadelphia chromosome causes fusion between ABL gene on chromosome9 and BCR gene on chromosome 22 due to chromosome transition. TheBCR-ABL fusion gene allows the production of a BCR-ABL fusion proteinwith abnormal tyrosine kinase activity. Activation of abnormal tyrosinekinase enzyme causes abnormal amplification of malignant cells, andthereby blood cancer occurs.

Gleevec® (imatinib) is a drug that competitively binds to the adenosinetriphosphate-binding site (ATP) in the BCR-ABL fusion protein andinhibits the enzymatic activity of the protein. However, in somepatients, mutations of the BCR-ABL gene lead to resistance to Gleevecand the disease get worse. As a patient group with the limits andresistance of Greeveck has emerged, second-generation (nilotinib) andthird-generation (dasatinib) tyrosine kinase inhibitors are beingdeveloped, but these drugs also have the disadvantage that completetreatment is not achieved, and the possibilty of treatment success inacute patients has increased by about 30%. Accordingly, research for thetreatment of chronic myelogenous leukemia is continuously needed.

Chronic lymphocytic leukemia is a disease in which lymphocytes, a typeof white blood cells, grow and turn into tumors, and accordingly,overgrow in the bone marrow, thereby interfering with the production ofnormal blood cells. When the number of normal white blood cellsdecreases, the risk of infection increases, and when the number of redblood cells decreases, anemia occurs, and the number of platelets thatact as a hemostatic agent decreases, so that the time for stopping thebleeding also increases. Chronic lymphocytic leukemia is very rare inKorea, but it occurs most frequently in the United States. It oftenappears in men after the age of 50. The cause of the onset of chroniclymphocytic leukemia has not yet been clarified, and is not related tothe environment or occupation, but also to virus or radiationirradiation. However, in the case of direct line families with chroniclymphocytic leukemia, the possibility that will develop chroniclymphocytic leukemia or other lymphoproliferative diseases increases bythree times as compared with a general population. When there is afamily history, it occurs at about 10 years younger than when it is notso.

Standard methods for treating leukemia include chemotherapy,hematopoietic stem cell transplantation, radiation therapy, and thelike. In the case of chemotherapy, a method of using in combination withtwo or more anticancer agents is usually included. The idealchemotherapy is that anti-leukemia drugs do not suppress normalhematopoiesis, and should show selective effects only on leukemic cells,without causing other harmful side effects. However, most anti-leukemiadrugs can kill leukemia cells in close proximity to the ideal state tosome degree, but because it also inhibits normal hematopoiesis andcauses other harmful side effects, there is a limit to the treatment ofleukemia. In addition, drug-resistant leukemia cells are weak inantitumor effect and may cause side effects, so that sufficientchemotherapy may not be performed.

Hematopoietic stem cell transplantation (HSCT) goes beyond the area ofbone marrow transplantation (BMT) that used bone marrow in the past, andat present, it means transplantation using all forms of hematopoieticmother cells present in peripheral blood (PB) and cord blood (CB) as atransplant source. Hematopoietic stem cell transplantation is atreatment method in which cancer cells and the patient's ownhematopoietic stem cells are removed using anticancer chemotherapy andradiation therapy in hematopoietic patients, and then new hematopoieticmother cells are transplanted. It is becoming an effective and promisingtreatment tool in various fields such as refractory autoimmune disease,and solid cancer as well as leukemia, aplastic anemia, malignantlymphoma, represented by leukemia, deviating from the limited concept ofearly bone marrow transplantation. However, until now, it is a treatmentmethod with a high incidence of complications due to high-dosechemotherapy treatment and graft-versus-host disease that occur afterallograft.

Therefore, for effective cancer treatment, it is important to establishand apply a treatment plan suitable for each cancer patient usingvarious methods such as radiotherapy, surgery, and chemotherapy. Inaddition, it is also an important task given to the industry thatdevelops new therapeutic agents for the treatment of various forms ofcancers, such as solid tumors and hematologic cancers.

On the other hand, as a prior literature related to 1,2-naphthoquinonederivative compounds, Korean Unexamined Patent Application PublicationNos. 10-2015-0080423, 10-2015-0080425 and 10-2015-0080426 disclose a1,2-naphthoquinone derivative and its preparation method, However, thesepatents relate to a composition for the treatment of metabolic diseases,and they have not disclosed that the 1,2-naphthoquinone derivativecompound having the structure of the present invention has a therapeuticeffect on solid cancer, acute leukemia, and hematocarcinoma such aschronic leukemia.

PRIOR LITERATURE Patent Literature

(Patent Literature 1) Korean Laid-open Patent Publication No.10-2015-0080423 (entitled 1,2-naphthoquinone derivative and preparationmethod thereof, published on Jul. 9, 2015)

(Patent Literature 2) Korean Laid-open Patent Publication No.10-2015-0080425 (entitled 1,2-naphthoquinone derivative and preparationmethod thereof, published on Jul. 9, 2015)

(Patent Literature 3) Korean Laid-open Patent Publication No.10-2015-0080426 (entitled 1,2-naphthoquinone derivative and preparationmethod thereof, published on Jul. 9, 2015)

DETAILED DESCRIPTION OF THE INVENTION Technical Problem

It is an object of the present invention to provide a pharmaceuticalcomposition for the prevention or treatment of solid cancers or bloodcancers, including a 1,2-naphthoquinone derivative compound.

Technical Solution

The present invention relates to a pharmaceutical composition for theprevention or treatment of solid cancers or blood cancers, including, asan active ingredient, a 1,2-naphthoquinone derivative compoundrepresented by the following Chemical Formula 1 or a pharmaceuticallyacceptable salt thereof. In a preferred embodiment, the blood cancer isacute leukemia, chronic leukemia, drug-resistant chronic leukemia orrefractory acute leukemia.

The present invention relates to a pharmaceutical composition for theprevention or treatment of solid cancers, including, as an activeingredient, a 1,2-naphthoquinone derivative compound represented by thefollowing Chemical Formula 1 or a pharmaceutically acceptable saltthereof.

in the Chemical Formula 1,

R₁ and R₂ are each independently hydrogen, halogen, hydroxy, C₁-C₆alkoxy, C₁-C₆ alkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₂-C₁₀ heteroaryl,—NO₂, —NR′₁R′₂, —NR′₁(CO(O)R′₂), —NR′₁(C(O)NR′₁R′₂), —CO(O)R′₁,—C(O)NR′₁R′₂, —CN, —SO(O)R′₁, —SO(O)NR′₁R′₂, —NR′₁(SO(O)R′₂),—CSNR′₁R′₂, or R₁ and R₂ taken together may form a cyclic structure ofC₄-C₁₀ aryl or a cyclic structure of C₂-C₁₀ heteroaryl, wherein R′₁ andR′₂ are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, orC₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl, —(CR″₁R″₂)_(m)—C₄-C₁₀aryl, —(CR″₁R″₂)_(m)—C₄-C₁₀ heteroaryl or NR″₁R″₂, the R″₁ and R″₂ areeach independently hydrogen, C₁-C₃ alkyl, or R″₁ and R″₂ taken togethermay form a cyclic structure of C₄-C₁₀ aryl,

R₃, R₄ and R₅ are each independently hydrogen, halogen, hydroxy, C₁-C₆alkyl, C₂-C₁₀ alkene, C₁-C₆ alkoxy, C₃-C₈ cycloalkyl, C₂-C₈heterocycloalkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₈ heteroaryl, —(CR′₅R′₆)_(m)—NR′₃R′₄,—(CR′₅R′₆)_(m)—C₃-C₈ heterocycloalkyl, —(CR′₅R′₆)_(m)—OR′₃,—(CR′₅R′₆)_(m)(O)COR′₃, —CO(O)R′₃, —CONR′₃R′₄, —NR′₃R′₄, —NR′₃(C(O)R′₄),—SO(O)R′₃, —SO(O)NR′₃R′₄, —NR′₃(SO(O)R′₄), —CSNR′₃R′₄, —CH₂A where thecompound of Chemical Formula 1 is “A”, or -A where the compound ofChemical Formula 1 is “A”, wherein the R′₃ and R′₄ are eachindependently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl, —CO(O)R″₃, or, R′₃ and R′₄ takentogether may form a cyclic structure of C₂-C₁₀ heterocycloalkyl, or acyclic structure of C₁-C₁₀ heteroaryl, the R′₅ and R′₆ are eachindependently hydrogen or C₁-C₃ alkyl, the R″₃ is C₁-C₆ alkyl;

Q₁ and Q₂ are each independently CO, COR₆, or COR₇,

when Q₁ is CO and Q₂ is CO, Q₁ and Q₂ form a single bond,

when Q₁ is COR₆ and Q₂ is COR₇, Q₁ and Q₂ form a double bond,

the R₆ and R₇ are each independently hydrogen, C₁-C₁₀ alkoxy,C₁-C₆alkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₂-C₁₀ heteroaryl, —CO(O)R′₇,—C(O)NR′₇R′₈, —SO(O)R′₇, —SO(O)NR′₇R′₈, —SO₃R′₇, —PO₃R′₇, —CSNR′₇R′₈, orR₆ and R₇ taken together may form a cyclic structure of C₃-C₁₀heterocyclo alkyl, or a cyclic structure of C₃-C₁₀ heteroaryl, the R′₇and R′₈ are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl, —(CR″4R″₅)m′—C₄-C₁₀ aryl,the R″₄ and R″₅ are each independently hydrogen, C₁-C₃ alkoxy;

Q₁ is a cyclic structure of substituted or unsubstituted C₃-C₅heterocyclo alkyl, Q₂ is CO, or, when Q₁ is CO and Q₂ is a cyclicstructure of substituted or unsubstituted C₃-C₅ heterocyclo alkyl, Q₁and Q₂ form a single bond;

m and m′ are each independently an integer of 1 to 4;

the hetero atom is at least one selected from N, O and S;

X₁ to X₄ are each independently CH or N(R″₆), X₅ is N, X₆ is C, X₇ is N,wherein R″₆ is hydrogen or C₁-C₃ alkyl;

in the Chemical Formula, the notation

means a single bond or a double bond, the notation

means that a single bond or a bond may not be formed, the notation

means that the cyclic structure including it may or may not be aromatic;and

the substituted means being substituted with one or more substituentsselected from the group consisting of hydroxy, halogen element, C₁-C₁₀alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₁₀ alkoxy, C₁-C₁₀alkoxycarbonyl, C₃-C₈ cycloalkyl, C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl,and C₂-C₁₀ heteroaryl.

The term “alkyl” means a linear or branched hydrocarbon group with asingle bond, and may include, for example, C₁-C₁₀ alkyl, specificallyC₁-C₆ alkyl, more specifically methyl, ethyl, propyl, n-butyl, isobutyl,tert-butyl, 1-methylpropyl, and the like.

The term “alkoxy” means an oxygen group to which a linear or branchedsaturated hydrocarbon with a single bond is bonded, and may include, forexample, C₁-C₁₀ alkoxy, specifically C₁-C₆ alkoxy, more specificallymethoxy, ethoxy, propoxy, n-butoxy, tert-butoxy, 1-methylpropoxy, andthe like.

The term “cycloalkyl” means a saturated hydrocarbon ring group with asingle bond, and may include, for example, C₃-C₁₀ cycloalkyl dependingon the number of carbon atoms, specifically C₃-C₈ cycloalkyl, morespecifically cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and thelike.

The term “heterocycloalkyl” means a saturated hydrocarbon ring groupwith a single bond including one or more heteroatoms such as N, O, or Sin addition to carbon atoms as ring members. Depending on the number andtype of heteroatoms contained in the ring, and the number of carbonatoms, for example, the heterocycloalkyl includes C₂-C₈heterocycloalkyl, C₂-C₁₀ heterocycloalkyl, or C₂-C₅ heterocycloalkylcontaining one or more, specifically, one to three heteroatoms selectedfrom the group consisting of N, O and S, more specifically, aziridine,pyrrolidine, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl,tetrahydrofuranyl or tetrahydropyranyl, and the like.

The term “aryl” means an aromatic substituent containing at least onering having a shared pi-electron system, and includes monocyclic orfused ring polycyclic (i.e., rings that share pairs of adjacent carbonatoms) groups. For example, depending on the number of carbon atomscontained in the ring, the aryl is specifically C₄-C₁₀ aryl, morespecifically C₆-C₁₀ aryl, and still more specifically phenyl, naphthyl,and the like.

The term “heteroaryl” means an aromatic cyclic compound containing oneor more heteroatoms such as N, O, or S in addition to a carbon atom as aring member. For example, depending on the number and type ofheteroatoms contained in the ring, and the number of carbon atoms, theheteroaryl includes C₁-C₁₀ heteroaryl, more specifically, C₁-C₈heteroaryl, C₂-C₁₀ heteroaryl, or C₂-C₅ heteroaryl, containing one ormore, specifically one to three heteroatoms selected from the groupconsisting of N, O, and S.

Examples of the aryl or heteroaryl include phenyl, naphthyl, furanyl,pyranyl, oxazolyl, isoxazolyl, imidazole, pyridyl, pyrazinyl, pyrimidyl,pyridazinyl, oxadiazolyl, thiadiazolyl, tetrazolyl, triazinyl, triazyl,and the like, but are not limited only thereto.

The term “aryloxy” means a group in which any one carbon forming anaromatic substituent is bonded to oxygen. For example, when oxygen isbonded to a phenyl group, it can be expressed as —O—C₆H₅, —C₆H₄—O—.

In the present invention, the “substituent” may be at least one,preferably one to three, selected from the group consisting of halo,hydroxy, a cyano group, a nitro group, an unsubstituted or substitutedalkyl group, an unsubstituted or substituted alkenyl group, anunsubstituted or substituted alkynyl group, an unsubstituted orsubstituted alkoxy group, an unsubstituted or substituted alkoxycarbonylgroup, an unsubstituted or substituted cycloalkyl group, anunsubstituted or substituted heterocycloalkyl group, an unsubstituted orsubstituted aryl group, and an unsubstituted and substituted heteroarylgroup. Specifically, the substituent may be at least one selected fromthe group consisting of hydroxy, halo, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl,C₂-C₁₀ alkynyl, C₁-C₁₀ alkoxy, C₁-C₁₀ alkoxycarbonyl, C₃-C₈ cycloalkyl,C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl, and C₂-C₁₀ heteroaryl.

In addition, the prodrug containing the 1,2-naphthoquinone derivativecompound of the present invention is regarded as one type of compoundexcluding the case where Q₁ is CO and Q₂ is CO in the ChemicalFormula 1. As an example included in the prodrug, the followingcompounds and the like correspond thereto.

Preferably, it may be a pharmaceutical composition for the treatment ofsolid cancers or blood cancers, including, as an active ingredient, a1,2-naphthoquinone derivative compound represented by the ChemicalFormula 1 or a pharmaceutically acceptable salt thereof, wherein:

in the Chemical Formula 1,

R₃ is hydrogen, halogen, hydroxy, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₃-C₈cycloalkyl, (CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—OR′₃, —CO(O)R′₃,—CH₂A where the compound of Chemical Formula 1 is “A”, or -A where thecompound of Chemical Formula 1 is “A”, wherein R′₃ is hydrogen, C₁-C₆alkyl, C₃-C₈ cycloalkyl, C₄-C₁₀ aryl, —(CR′R′)_(m)—C₄-C₁₀ aryl,—(CR′R′)_(m)—C₄-C₁₀ aryloxy, —(CR′₅R′)_(m)—C₁-C₁₀ heteroaryl, —CO(O)R″₃,the R′₅ and R′₆ are each independently hydrogen, C₁-C₆ alkyl, the R″₃ isC₁-C₆ alkyl;

R₄ is hydrogen, halogen, hydroxy, C₁-C₆ alkyl, C₂-C₁₀ alkene, C₁-C₆alkoxy, C₃-C₈ cycloalkyl, C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl, C₄-C₁₀aryloxy, C₁-C₈ heteroaryl, —(CR′5R′₆)_(m)—C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy, —(CR′₅R′₆)_(m)—C₁-C₈ heteroaryl,—(CR′₅R′₆)_(m)—NR′₃R′₄, —(CR′₅R′₆)_(m)—C₃-C₈ heterocycloalkyl,—(CR′₅R′₆)_(m)—OR′₃, —(CR′₅R′₆)_(m)(O)COR′₃, —CO(O)R′₃, —CONR′₃R′₄,—NR′₃R′₄, —NR′₃(C(O)R′₄), -A where the compound of Chemical Formula 1 is“A”, wherein R′₃ and R′₄ are each independently hydrogen, C₁-C₆ alkyl,C₃-C₈ cycloalkyl, C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy, —(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl,—CO(O)R″₃, or R′₃ and R′₄ taken together may form a cyclic structure ofC₂-C₁₀ heterocycloalkyl, or a cyclic structure of C₁-C₁₀ heteroaryl, theR′₅ and R′₆ are each independently hydrogen or C₁-C₃ alkyl, the R″₃ isC₁-C₆ alkyl;

R₅ is hydrogen, halogen, hydroxy, C₁-C₆ alkyl, C₂-C₁₀ alkene, C₁-C₆alkoxy, C₃-C₈ cycloalkyl, C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl, C₄-C₁₀aryloxy, C₁-C₈ heteroaryl, —(CR′5R′₆)_(m)—C₄-C₁₀ aryl,—(CR′5R′₆)_(m)—C₄-C₁₀ aryloxy, —(CR′₅R′₆)_(m)—C₁-C₈ heteroaryl,—(CR′₅R′₆)_(m)—NR′₃R′₄, —(CR′₅R′₆)_(m)—C₃-C₈ heterocycloalkyl,—(CR′₅R′₆)_(m)—OR′₃, —(CR′₅R′₆)_(m)(O)COR′₃, —CO(O)R′₃, —CONR′₃R′₄,—NR′₃R′₄, —NR′₃(C(O)R′₄), —CH₂A where the compound of Chemical Formula 1is “A”, wherein R′₃ and R′₄ are each independently hydrogen, C₁-C₆alkyl, C₃-C₈ cycloalkyl, C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy, —(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl,—CO(O)R″₃, or R′₃ and R′₄ taken together may form a cyclic structure ofC₂-C₁₀ heterocycloalkyl, or a cyclic structure of C₁-C₁₀ heteroaryl, theR′₅ and R′₆ are each independently hydrogen or C₁-C₃ alkyl, and the R″₃is C₁-C₆ alkyl;

Further, a more specific example of the compound of the Chemical Formula1 is as follows.

In addition, the present invention relates to a pharmaceutical forpreventing or treating solid cancers or blood cancers comprising a1,2-naphthoquinone derivative compound or a pharmaceutically acceptablesalt thereof as an active ingredient, characterized in that the compoundis one of the compounds represented below.

In the present invention, the solid cancer may be one or more cancersselected from the group consisting of gastric cancer, liver cancer,colon cancer, breast cancer, lung cancer, non-small cell lung cancer,pancreatic cancer, skin cancer, head and neck cancer, uterine cancer,ovarian cancer, colorectal cancer, small intestinal cancer, rectalcancer, prostate cancer, esophageal cancer, malignant lymphs, bladdercancer, gallbladder cancer, kidney cancer, and brain tumor and the like.Preferably, the cancer may be one or more cancers selected from thegroup consisting of lung cancer, uterine cancer, liver cancer, andbreast cancer.

Among the blood cancers, the acute leukemia may be one or more cancersselected from the group consisting of acute myeloid leukemia and acutelymphocytic leukemia.

Further, the blood cancer may be drug-resistant or refractory leukemiahaving resistance to existing anticancer drugs. Specifically, suchdrug-resistant or refractory leukemia is drug-resistant refractory acuteleukemia having resistance to idarubicin or cytarabine, which is atherapeutic agent for acute leukemia, or a drug-resistant chronicleukemia having resistance to imatinib, which is a therapeutic agent forchronic leukemia.

A pharmaceutically acceptable salt of the 1,2-naphthoquinone derivativecompound of the present invention may include addition salts formed byinorganic acids such as hydrochloride, sulfate, phosphate, hydrobromide,hydroiodide, nitrate, pyrosulfate, or metaphosphate, addition saltsformed by organic acids such as citrate, oxalate, benzoate, acetate,trifluoroacetate, propionate, succinate, fumarate, lactate, maleate,tartrate, glutarate, or sulfonate, or metal salts such as lithium salt,sodium salt, potassium salt, magnesium salt and calcium salt, but is notlimited thereto.

The pharmaceutical composition according to the present invention can beformulated into a suitable form together with a commonly usedpharmaceutically acceptable carrier. The “pharmaceutically acceptable”refers to being physiologically acceptable, and not usually causing anallergic reaction or a similar reaction such as gastrointestinaldisorders and dizziness when administered to humans. Further, thepharmaceutical composition of the present invention may be used afterbeing formulated into an oral preparation, such as powders, granules,tablets, capsules, suspensions, emulsions, syrups, and aerosols, etc.,and a parental preparation, such as epidermal formulations,suppositories, or sterile injection solutions, in accordance with aconventional method.

Examples of carriers, excipients and diluents that can be included inthe composition, may include lactose, dextrose, sucrose, sorbitol,mannitol, xylitol, erythritol, maltitol, starch, arabic gum, alginate,gelatin, calcium phosphate, calcium silicate, cellulose,methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone,water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesiumstearate, and mineral oil, but are not limited thereto. When formulatedinto a preparation, a diluting agent or an excipient, such ascommonly-used fillers, stabilizing agents, binding agents,disintegrating agents, and surfactants can be used. Solid preparationsfor oral administration include tablets, pills, powders, granules,capsules, and the like, and these solid preparations may be prepared bymixing the compound of the present invention with at least oneexcipient, for example, starch, microcrystalline cellulose, sucrose,lactose, low-substituted hydroxypropyl cellulose, hypromellose or thelike. In addition to the simple excipient, a lubricant such as magnesiumstearate and talc are also used. Liquid preparations for oraladministration include a suspension, a liquid for internal use, anemulsion, a syrup, etc. In addition to a commonly used simple diluentsuch as water and liquid paraffin, various excipients such as ahumectant, a sweetener, an aromatic, a preservative, etc. may also becontained. Formulations for parenteral administration include asterilized aqueous solution, a non-aqueous solution, a suspension, anemulsion, a lyophilized formulation and a suppository. The non-aqueoussolution or suspension may contain propylene glycol, polyethyleneglycol, a vegetable oil such as olive oil, an injectable ester such asethyl oleate, etc. As a base of the suppository, witepsol, macrogol,tween 61, cocoa butter, laurin butter, glycerogelatin, etc. may be used.In order to formulate the formulation for parenteral administration, the1,2-naphthoquinone derivative compound of Chemical Formula 1 or apharmaceutically acceptable salt thereof may be mixed in water togetherwith sterilized and/or contain adjuvants such as preservatives,stabilizers, auxiliary agents such as wettable powder or emulsifyingaccelerators, salt for controlling osmotic pressure and/or buffers andthe like, and other therapeutically useful substances, to prepare asolution or suspension, which is then manufactured in the form of anampoule or vial unit administration.

The pharmaceutical composition including the compound of ChemicalFormula 1 disclosed herein as an active ingredient may be administeredto mammals such as mice, livestock, and humans by various routes for theprevention or treatment of solid cancers or blood cancers. All modes ofadministration can be predicted, and for example, it can be administeredby oral, rectal or intravenous, intramuscular, subcutaneous, endometrialor cerebrovascular injection. The dosage is varied depending on the age,sex, weight of the subject to be treated, the specific disease orpathological condition to be treated, the severity of the disease orpathological condition, the duration of administration, the route ofadministration, the drug absorption, distribution and excretion rate,the types of other drugs used, the judgment of prescriber, and the like.Dosage determination based on such factors is within the standards ofthose skilled in the art, and the dosage generally ranges from 0.01mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage is 1mg/kg/day to 500 mg/kg/day. It can be administered once a day or inseveral divided doses. The dosage does not limit the scope of thepresent invention in any way.

Advantageous Effects

The present invention relates to a pharmaceutical composition for theprevention or treatment of solid cancers or blood cancers such as acuteleukemia or chronic leukemia, the pharmaceutical composition including,as an active ingredient, a 1,2-naphthoquinone derivative compound or apharmaceutically acceptable salt thereof. When solid cancer, acuteleukemia, and chronic leukemia, and drug-resistant/refractory leukemiacell lines are treated with the 1,2-naphthoquinone derivative compound,it is excellent in the effect of killing cells and thus, can be usefullyused as a pharmaceutical composition for preventing or treating thecarcinoma.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the cell viability of HL60 (FIG. 1A) or U937(FIG. 1B) cells when treating with the compound 1 of the presentinvention.

FIG. 2 is a result of confirming the apoptosis or cell necrosis of KG1acells by FACS (FIG. 2A) and a result of confirming them by Western blot(FIG. 2B), when treating with the compound 1 of the present invention.

FIG. 3 is a result of confirming the apoptosis or cell necrosis of HL60cells by FACS (FIG. 3A) and a result of confirming them by Western blot(FIG. 3B), when treating with the compound 1 of the present invention.

FIG. 4 is a result of confirming the proliferation inhibitory effect ofmonocytic cells in peripheral blood (FIG. 4A) or spleen (FIGS. 4B and4C), when treating with the compound 1 of the present invention in amouse model of an acute leukemia in which FLT3-ITD is overexpressed.

FIG. 5 is a result of confirming the expression level of the BCR-ABLfusion gene by Western blot by treating K562 cells with the compound 1of the present invention according to concentration.

FIG. 6 is a graph showing the cell viability of A549 (lung carcinoma)and Hela (cervix adenocarcinoma) cells by treating A549 (lung carcinoma)and Hela (cervix adenocarcinoma) with the compounds 1, 10 and 72 of thepresent invention.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Hereinafter, preferred examples of the present invention will bedescribed in detail. However, the present invention is not limited tothe examples described herein, and can also be embodied in other forms.Rather, the content presented herein will be thorough and complete, andwill fully convey the scope of the present invention to those skilled inthe art.

Example 1. Synthesis of 1,2-Naphthoquinone Derivative Compound

As the 1,2-naphthoquinone derivative compound of the present inventionused for confirming the therapeutic effect on acute leukemia, thecompounds 1 to 98 and 175 to 190 were synthesized with reference to themethod for synthesizing the compounds disclosed in Korean PatentApplication Nos. 10-2014-0193184, 10-2014-0193306, 10-2014-0193370,10-2015-0043050, and 10-2015-0043068. In addition, among the compoundsprepared through the above process, the physicochemical properties forthe compound 98 are shown in Table 1 below.

TABLE 1 Compound No. ¹H NMR data

¹H NMR (300 MHz, DMSO) δ 7.95-7.86 (m, 2H), 7.70 (t, J = 7.5 Hz, 1H),7.45 (t, J = 7.5 Hz, 1H), 4.99 (d, J = 5.7 Hz, 2H), 4.5 (d, J = 6.0 Hz,2H), 1.75 (s, 3H)

Example 2. Preparation of Acute Leukemia Cell Line,Measurement/Confirmation of Cell Viability, Apoptosis or Cell NecrosisThereof Example 2-1. Preparation of Acute Leukemia Cell Line

The causative factor of acute leukemia has been found to be verydiverse. The constructed cell lines were also diverse. Thus, in order toconfirm the applicability to a wide range of therapeutic agents that arenot therapeutic agents for specific types of acute leukemia, in thepresent invention, KG1α cells constructed by selecting only cells havinga stem cell phenotype from KG1 cells, which are cell lines obtained froma 59-year-old man with acute myelogenous leukemia, were used. HL60cells, which are cell lines obtained from a 36-year-old woman with acutepromyelocytic leukemia, and U937 cells, which are cell lines obtainedfrom a 37-year-old man with histiocytic lymphoma, were used.

KG1α and HL60 cells were cultured in IMDM medium containing 20% FBS(fetal bovine serum), and U937 cells were cultured in RPMI 1640 mediumcontaining 10% FBS. All cells were cultured in an incubator underconditions of 37° C. and 5% CO₂, and subcultured once every 2 or 3 daysand used in the experiment.

Example 2-2. Measurement of Cell Viability in KG1α Cells-WST Assay

KG1α cells are cells resistant to idarubicin and cytarabine, which arethe therapeutic agents for acute myeloid leukemia, and are refractoryAML cells. KG1α cells cultured in Example 2-1 were cultured into a96-well plate at 1×10⁵ cells/well, and then cultured in an incubator at37° C. and 5% CO₂ for 16 hours or more for the stabilization of cells.Subsequently, among the compounds synthesized in Example 1, thecompounds 1, 10 and 72 were treated at a concentration of 0.1 to 3 μM,respectively. as shown in Table 2 below, and then cultured for 24 hours.At this time, DMSO was used as a control group. After 24 hours, eachcell was treated with 10 μl of WST solution, reacted for 2 hours, andthen absorbance was measured at 450 nm with a multi-scan machine, andthe cell viability of KG1α cells was confirmed as shown in Table 4below.

TABLE 2 IC₅₀ Condition (μM)  

0.5

0.5

0.7

Table 2 shows the cell viability of KG1α cells when treated withcompounds 1, 10 and 72 of the present invention as compared with thecontrol group. The IC₅₀ value appeared as 0.5 to 1 μM, confirming thatthe effect of killing acute myeloid cells was excellent.

Example 2-3. Measurement of Cell Viability of HL60 Cells or U937 Cells

The HL60 cells or U937 cells cultured in Example 2-1 were inoculatedinto a 96-well plate at 1×10⁵ cells/well, and then for the stabilizationof cells, the cells were cultured in an incubator at 37° C. and 5% CO₂for 16 hours or more. Subsequently, the compound 1 synthesized inExample 1 was treated at concentrations of 0.1, 0.5 and 1 μM,respectively, and reacted for 24 hours. At this time, DMSO anddecitabine having a concentration of 0.1 or 1 μM was used as a controlgroup. After 24 hours, each cell was treated with 10 μl of WST-1solution, and reacted for 2 hours. The absorbance was then measured at450 nm with a multi-scan machine. The cell viability of HL60 cells isshown in FIG. 1A, and the cell viability results of U937 cells are shownin FIG. 1B.

Referring to the results of FIG. 1 , when HL-60 cells or U937 cells wastreated with the compound 1 of the present invention, it was confirmedthat it exhibited concentration-dependent cell killing effects in bothHL-60 cells or U937 cells. However, when HL-60 cells or U937 cells weretreated with a positive control decitabine, it exhibitedconcentration-dependent cell killing effects in HL60 cells, but in U937cells, the cell killing effect was remarkably low. Therefore, it wasconfirmed that the 1,2-naphthoquinone derivative compound of the presentinvention can be usefully used as a composition for treating a widerange of acute leukemias, rather than a therapeutic agent for a specifictype of acute leukemia.

Example 2-4. Confirmation of Apoptosis or Cell Necrosis in AcuteLeukemia Cells

Cell death was typically known as apoptosis or cell necrosis. Thus, inorder to confirm whether the death of acute leukemia cells induced bythe compound 1 of the present invention was due to apoptosis or cellnecrosis, fluorescence activated cell sorting (FACS) and western blotwere performed.

First, in order to conduct FACS, KG1α cells or HL60 cells cultured inExample 1-2 were inoculated in a 6-well plate at 1×10⁷ cells/well, andfor the stabilization of cells, the cells were cultured in an incubatorat 37° C. and 5% CO₂ for 16 hours or more. Subsequently, the compound 1was treated at concentrations of 0, 0.1, 0.5, 1 and 2 μM, respectively,and cultured for 24 hours, and then the cells were recovered and washedwith cold PBS. Only 1×10⁶ cells were taken and resuspended with PBS, andthen stained with Annexin-V (green fluorescence), which increases duringapoptosis, and Pi (propidium iodide, red fluorescence), which reactsduring apoptosis. The degree of the two indicators were quantified byFACS, the FACS results for KG1α cells are shown in FIG. 2A, and the FACSresults for HL60 cells are shown in FIG. 3A.

Next, in order to perform western blot, KG1α cells or HL60 cellscultured in Example 2-1 were inoculated into a 6-well plate at 1×10⁷cells/well. In order to stabilize the cells, the cells were cultured for16 hours or more in an incubator at 37° C. and 5% CO₂. Then, thecompound 1 was treated at concentrations of 0, 0.1, 0.5, 1 and 2 μM,respectively, and cultured for 24 hours, and then only cells wererecovered. The protein extraction buffer was added to the cells obtainedin the above process, and then reacted on ice for 30 minutes to breakthe cell membrane and centrifuged to take only the supernatant, and thenextract the protein. SDS-PAGE was performed using the extracted protein,and then transferred to the nitrocellular membrane. Then, after reactinga specific antibody related to apoptosis, the amount of the protein wasconfirmed using the ECL buffer, and the results of Western blotting ofKG1α cells are shown in FIG. 2B, and the results of Western blotting ofHL60 cells are shown in FIG. 3B.

Referring to the results of FIGS. 2 and 3 , when U937 cells or A549cells were treated with the compound 1 of the present invention, in KG1αcells, acute leukemia cells died due to apoptosis, but in HL60 cells,both apoptosis and cell necrosis occurred, confirming that acuteleukemia cells died.

Example 3. Confirmation of Changes in Monocyte Cells in Acute LeukemiaModel

In order to confirm the change in the expression of the FLT3-ITD gene byCompound 1 of the present invention, peripheral blood and spleen werecollected after 8 weeks from the animals of an experimental group inwhich Compound 1 of the present invention was mixed and administered ina diet at a concentration of 120 mg/kg using a mouse (jackson lab) thatsystemically overexpressed FLT3-ITD, and a control group to which only asolvent was administered (untreated group).

In the case of peripheral blood, cold 1×PBS was added to a small amountof blood, centrifuged, washed, to which RBC lysis buffer was added andreacted for 5 minutes at room temperature to remove red blood cells.After adding cold 1×PBS again, the pellet from which the supernatant hadbeen removed by centrifugation was resuspended in 1×PBS containing 1%FBS. Only 1×10⁶ cells were taken, and Gr-1 and Mac-1 antibodies, whichwere specifically expressed in granulocytes, and CD3 antibodyspecifically expressed in monocytes, were simultaneously immuno-stained.The antibodies were reacted on ice at a ratio of 1:200 for 20 minutes,washed with 1×PBS, then resuspended with 1×PBS containing 1% FBS, andquantified by FACS, and shown in FIG. 4A.

In the case of the spleen, the cells were crushed on a mesh having a 40μm hole, separated into single cells, and then washed with cold 1×PBS towhich RBC lysis buffer was added and reacted at room temperature for 5minutes to remove red blood cells. After washing again with cold 1×PBS,the pellet was resuspended in 1×PBS containing 1% FBS. Only 1×10⁶ cellswere taken, and Gr-1 and Mac-1 antibodies, which were specificallyexpressed in granulocytes, and CD3 antibody specifically expressed inmonocytes, were simultaneously immuno-stained. The antibodies werereacted on ice at a ratio of 1:200 for 20 minutes, washed with 1×PBS,resuspended in 1×PBS containing 1% FBS, and quantified by FACS, andshown in FIGS. 4B and 4C.

Referring to the results of FIG. 4 , when the compound 1 of the presentinvention was treated in an acute myelogenous leukemia mouse model inwhich FLT3-ITD was overexpressed, it was confirmed that it had anexcellent effect of suppressing the proliferation of monocyte cells(acute leukemia) in both peripheral blood and spleen, and thus had aneffect of improving acute myeloid leukemia.

Example 4. Preparation of Chronic Leukemia Cell Line and Measurement ofCell Viability Thereof Example 4-1. Preparation of Chronic Leukemia CellLines

K562 cells obtained from a 53-year-old woman with chronic myeloidleukemia were cultured in an incubator under conditions of 37° C. and 5%CO₂ using RPMI medium containing 10% FBS (fetal bovine serum), andsubcultured once every two days and used in the experiment.

Example 4-2. Measurement of Cell Viability in Chronic Leukemia Cells-WSTAssay

The K562 cells cultured in Example 4-1 were inoculated into a 96-wellplate at 1×10⁵ cells/well, and then, for the stabilization of cells, thecells were cultured in an incubator at 37° C. and 5% CO₂ for 16 hours ormore. Subsequently, among the compounds synthesized in Example 1, thecompounds 1, 10, and 72 and 192 were treated at a concentration of 0.1to 5 μM and then cultured for 4 hours. At this time, DMSO was used as acontrol group. After 4 hours of incubation, 10 μl of WST solution wasadded to each cell and then reacted for 2 hours. The absorbance was thenmeasured at 450 nm with a multi-scan machine, and the cell viability ofK562 cells is shown in Table 3 below.

TABLE 3 IC₅₀ Condition (μM)  

1.6

0.9

1.4

Referring to Table 3 above, when K562 cells were treated with thecompounds 1, 10, and 72 of the present invention, it was confirmed thatthe IC₅₀ value appeared as 1 to 2 μM, confirming that the effect ofkilling chronic myelogenous leukemia cells was excellent, as comparedwith the control group.

Example 5. Confirmation of Expression Level of BCR-ABL Fusion Gene inChronic Myelogenous Leukemia Cells

The BCR-ABL fusion gene, existing only in chronic myelogenous leukemia,generated and transmitted continuous cell growth signals to induce thegrowth of cancer cells. Thus, after chronic myeloid leukemia cells weretreated with the compound of the present invention, western blot wasused to confirm whether the expression level of the BCR-ABL fusion genewas reduced.

First, the K562 cells cultured in Example 4-1 were inoculated into a 60mm plate at 5×10⁶ cells/well, and then for the stabilization of cells,the cells were cultured in an incubator at 37° C. and 5% CO₂ for 16hours or more. The, the compound 1 was treated at concentrations of 0.5,1, 1.5, 2 and 2.5 μM, respectively, and cultured for 6 hours, and thenthe cells were recovered. A protein extraction buffer (RIPA buffer) wasadded to the cells obtained in the above process, and reacted on ice for30 minutes to break the cell membrane, which was centrifuged to remove asupernatant, and then the protein was extracted. The extracted proteinwas developed by electrophoresis using SDS-PAGE, and then transferred tothe PVDF membrane. Then, the antibody related to the BCR-ABL fusion genewas reacted, and then the amount of the protein was confirmed using theECL buffer, and Western blot results for the K562 cell line are shown inFIG. 5 .

Referring to the results of FIG. 5 , when K562 cells were treated withthe compound 1 of the present invention, it was confirmed that theexpression level of the BCR-ABL fusion gene, which plays the mostimportant role in the onset of chronic myelogenous leukemia, wasdecreased in a concentration-dependent manner. In addition, it can beseen that due to the decrease in the expression of the BCR-ABL fusiongene, the expression of phospho-bcr-abl and phospho-stat5, whichindicates the activity of the BCR-ABL fusion gene (a signal that inducescell growth), was also decreased.

However, the expression level of the BCR-ABL fusion gene, which is acancer-causing gene, decreased, whereas in the case of c-abl proteinpresent in normal blood cells, the expression level was not affected.Thus, it was confirmed that the compound of the present invention doesnot affect the c-abl protein expressed in normal cells, and thecomposition selectively reduces only the BCR-ABL fusion protein which isa cancer-causing gene existing only in chronic myelogenous leukemia.

Example 6. Preparation of Solid Cancer Cell Lines and Measurement ofCell Viability Thereof Example 6-1. Preparation of Solid Cancer CellLines

To confirm the possibility of use as a therapeutic agent for solidcancer, A549 cell line obtained from a 58-year-old men with lungcarcinoma, Hela cell line obtained from a 31-year-old women with cervixadenocarcinoma, HepG2 cell line obtained from a 15-year-old boy withhepatocellular carcinoma, MCF7 cell line obtained from a 69-year-oldwoman with breast adenocarcinoma, and Beas-2B cell line obtained from anormal lung for comparison with solid cancer cell line were used.

The A549 (lung carcinoma), Hela (cervix adenocarcinoma), HepG2(hepatocellular carcinoma), MCF7 (breast carcinoma), and Beas-2B (normallung) cell lines were used in DMEM medium containing 10% FBS,subcultured once every 2 days in an incubator under conditions of 37° C.and 5% CO₂ and used in the experiment.

Example 6-2. Measurement of Cell Viability in Solid Cancer Cells-WSTAssay

A549 (lung carcinoma), Hela (cervix adenocarcinoma), HepG2(hepatocellular carcinoma), MCF7 (breast carcinoma), and Beas-2B (normallung) cells cultured in Example 6-1 were inoculated into a 96-wellplated at 1×10⁴ cells/well, and then for the stabilization of cells, thecells were cultured in an incubator at 37° C. and 5% CO₂ for 16 hours ormore. Subsequently, among the compounds synthesized in Example 1, thecompounds 1, 10 and 72 were treated at a concentration of 1 to 30 μM,and then cultured for 6 hours. At this time, DMSO and β-lapachone wereused as the control group. After 4 hours of incubation, 10 μl of WSTsolution was added to each cell, the reaction was carried out for 2hours, and the absorbance was measured at 450 nm with a multi-scanmachine. The cell viabilities of A549 (lung carcinoma), Hela (cervixadenocarcinoma), HepG2 (hepatocellular carcinoma) and MCF7 (breastcarcinoma) cells are shown in Table 4 and FIG. 6 below.

TABLE 4 IC₅₀ (μM) Condition A549 Hela HepG2 MCF7 Positive control group5.0 4.8 12.3 13.2 (beta-rapacon)

5.1 4.6 10.1 14.4

5.5 7.7 12.1 14.1

6.5 9.1 12.6 12.5

Referring to Table 4 and FIG. 6 , when A549 (lung carcinoma), Hela(cervix adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breastcarcinoma) and Beas-2B (normal lung) cells were treated with thecompounds 1, 10 and 72 of the present invention for 6 hours, it wasconfirmed that they exhibited a killing effect similar to β-lapachone,and thus, the compounds of the present invention had a therapeuticeffect on solid cancer, as compared with the control group.

Further, although not shown in Table 4, when Beas-2B which is a normallung cell line was treated with the compounds of the present invention,the average cell viability was 80% or more, but when treating A549 whichis a lung carcinoma, it exhibited the cell viability of about 50%. Thus,since the 1,2-naphthoquinone derivative compound of the presentinvention did not exhibit cytotoxicity to normal cells but specificallyexhibited apoptosis effect on cancer cells, it can be confirmed that itcan be usefully used as a composition for the treatment of various solidcancers.

Example 7. Toxicity Experiment

This experiment was conducted to examine the acute toxicity to theanimal body acutely (within 24 hours) when the compound 1 of the presentinvention was ingested in excess in a short period of time, and todetermine the mortality rate. 20 C₅₇BL/6 mice, which are common mice,were prepared, and ten mice were assigned to each group. Only 0.1% SLS(sodium lauryl sulfate) was administered to the control group, and thecompound 1 was orally administered to the experimental group at aconcentration of 120 mg/kg, respectively. As a result of examining eachmortality rate 24 hours after administration, both the control group andthe experimental group to which the compound 1 was administeredsurvived.

Formulation Example 1. Preparation of Pharmaceutical FormulationContaining the Compound of the Present Invention Formulation Example1-1. Preparation of Powder

2 g of the compound 1 of the present invention and 1 g of lactose weremixed and filled in an airtight cloth to prepare a powder.

Formulation Example 1-2. Preparation of Tablets

100 mg of the compound 1 of the present invention, 100 mg ofmicrocrystalline cellulose, 60 mg of lactose hydrate, 20 mg oflow-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearatewere mixed, and then the mixture was tableted according to aconventional tablet preparation method to prepare tablets.

Formulation Example 1-3. Preparation of Capsules

100 mg of the compound 1 of the present invention, 100 mg ofmicrocrystalline cellulose, 60 mg of lactose hydrate, 20 mg oflow-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearatewere mixed, and then the above ingredients were mixed according to aconventional capsule preparation method and filled into gelatin capsulesto prepare capsules.

Formulation Example 1-4. Preparation of Injections

10 mg of the compound 1 of the present invention, an appropriate amountof injectable sterilized distilled water and an appropriate amount of apH adjuster were mixed, and then injections were prepared in the amountof the above components per ampoule (2 ml) according to a conventionalmethod for preparing injections.

The invention claimed is:
 1. A method for treating solid cancers orblood cancers, comprising, administering to a subject a therapeuticallyeffective amount of a pharmaceutical composition comprising, as anactive ingredient, a 1,2-naphthoquinone derivative compound representedby the following Chemical Formula 1 or a pharmaceutically acceptablesalt thereof:

wherein: R₁ and R₂ are each independently hydrogen, halogen, hydroxy,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₂-C₁₀heteroaryl, —NO₂, —NR′₁R′₂, —NR′₁(CO(O)R′₂), —NR′₁(C(O)NR′₁R′₂),—CO(O)R′₁, —C(O)NR′₁R′₂, —CN, —SO(O)R′₁, —SO(O)NR′₁R′₂, —NR′₁(SO(O)R′₂),—CSNR′₁R′₂, or R₁ and R₂ taken together form a cyclic structure ofC₄-C₁₀ aryl or a cyclic structure of C₂-C₁₀ heteroaryl, wherein R′₁ andR′₂ are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, orC₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl, —(CR″₁R″₂)_(m)—C₄-C₁₀aryl, —(CR″₁R″₂)_(m)—C₄-C₁₀ heteroaryl or NR″₁R″₂, the R″₁ and R″₂ areeach independently hydrogen, C₁-C₃ alkyl, or R″₁ and R″₂ taken togetherform a cyclic structure of C₄-C₁₀ aryl; R₃, R₄ and R₅ are eachindependently hydrogen, halogen, hydroxy, C₁-C₆ alkyl, C₂-C₁₀ alkene,C₁-C₆ alkoxy, C₃-C₈ cycloalkyl, C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl,C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy, —(CR′₅R′₆)_(m)—C₁-C₈ heteroaryl,—(CR′₅R′₆)_(m)—NR′₃R′₄, —(CR′₅R′₆)_(m)—C₃-C₈ heterocycloalkyl,—(CR′₅R′₆)_(m)—OR′₃, —(CR′₅R′₆)_(m)(O)COR′₃, —CO(O)R′₃, —CONR′₃R′₄,—NR′₃R′₄, —NR′₃(C(O)R′₄), —SO(O)R′₃, —SO(O)NR′₃R′₄, —NR′₃(SO(O)R′₄),—CSNR′₃R′₄, —CH₂A where the compound of Chemical Formula 1 is “A”, or -Awhere the compound of Chemical Formula 1 is “A”, wherein the R′₃ and R′₄are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₄-C₁₀aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl, —CO(O)R″₃, or, R′₃ and R′₄ takentogether form a cyclic structure of C₂-C₁₀ heterocycloalkyl, or a cyclicstructure of C₁-C₁₀ heteroaryl, the R′₅ and R′₆ are each independentlyhydrogen or C₁-C₃ alkyl, the R″₃ is C₁-C₆ akyl; Q₁ and Q₂ are eachindependently CO, COR₆, or COR₇, when Q₁ is CO and Q₂ is CO, Q₁ and Q₂form a single bond, when Q₁ is COR₆ and Q₂ is COR₇, Q₁ and Q₂ form adouble bond; R₆ and R₇ are each independently hydrogen, C₁-C₁₀ alkoxy,C₁-C₆ alkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₂-C₁₀ heteroaryl, —CO(O)R′₇,—C(O)NR′₇R′₈, —SO(O)R′₇, —SO(O)NR′₇R′₈, —SO₃R′₇, —PO₃R′₇, —CSNR′₇R′₈, orR₆ and R₇ taken together form a cyclic structure of C₃-C₁₀heterocycloalkyl, or a cyclic structure of C₃-C₁₀ heteroaryl, the R′₇and R′₈ are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl, —(CR″₄R″₅)_(m)′—C₄-C₁₀aryl, the R″₄ and R″₅ are each independently hydrogen, C₁-C₃ alkoxy; mand m′ are each independently an integer of 1 to 4; the hetero atom isat least one selected from N, O and S; X₁ to X₄ are each independentlyCH or N(R″₆); X₅ is N, X₆ is C, X₇ is N, wherein R″₆ is hydrogen orC₁-C₃ alkyl; in the Chemical Formula, the notation

means a single bond or a double bond, the notation

means that a single bond or a bond may not be formed, the notation

means that the cyclic structure may be or may not be aromatic; and thesubstituted means being substituted with one or more substituentsselected from the group consisting of hydroxy, halogen element, C₁-C₁₀alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₁₀ alkoxy, C₁-C₁₀alkoxycarbonyl, C₃-C₈ cycloalkyl, C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl,and C₂-C₁₀ heteroaryl; wherein the blood cancer is selected from thegroup consisting of acute leukemia, chronic leukemia and drug-resistantchronic leukemia or refractory acute leukemia; the chronic leukemia isselected from the group consisting of chronic myelogenous leukemia orchronic lymphocytic leukemia; the acute leukemia is selected from thegroup consisting of acute myelogenous leukemia or acute lymphocyticleukemia; and the solid cancer is selected from the group consisting oflung cancer, uterine cancer, liver cancer and breast cancer.
 2. Themethod according to claim 1, wherein the pharmaceutical compositioncomprises, as an active ingredient, a 1,2-naphthoquinone derivativecompound represented by the Chemical Formula 1 or a pharmaceuticallyacceptable salt thereof, wherein: R₃ is hydrogen, halogen, hydroxy,C₁-C₆ alkyl, C₁-C₆ alkoxy, C₃-C₈ cycloalkyl, (CR′₅R′₆)_(m)—C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—OR′₃, —CO(O)R′₃, —CH₂A where the compound of ChemicalFormula 1 is “A”, or -A where the compound of Chemical Formula 1 is “A”,wherein R′₃ is hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₄-C₁₀ aryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl, —CO(O)R″₃, the R′₅ and R′₆ are eachindependently hydrogen, C₁-C₆ alkyl, the R″₃ is C₁-C₆ akyl; R₄ ishydrogen, halogen, hydroxy, C₁-C₆ alkyl, C₂-C₁₀ alkene, C₁-C₆ alkoxy,C₃-C₈ cycloalkyl, C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy,C₁-C₈ heteroaryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀aryloxy, —(CR′₅R′₆)_(m)—C₁-C₈ heteroaryl, —(CR′₅R′₆)_(m)—NR′₃R′₄,—(CR′₅R′₆)_(m)—C₃-C₈ heterocycloalkyl, —(CR′₅R′₆)_(m)—OR′₃,—(CR′₅R′₆)_(m)(O)COR′₃, —CO(O)R′₃, —CONR′₃R′₄, —NR′₃R′₄, —NR′₃(C(O)R′₄),-A where the compound of Chemical Formula 1 is “A”, wherein R′₃ and R′₄are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₄-C₁₀aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl, —CO(O)R″₃, or R′₃ and R′₄ takentogether form a cyclic structure of C₂-C₁₀ heterocycloalkyl, or a cyclicstructure of C₁-C₁₀ heteroaryl, the R′₅ and R′₆ are each independentlyhydrogen or C₁-C₃ alkyl, the R″₃ is C₁-C₆ akyl; R₅ is hydrogen, halogen,hydroxy, C₁-C₆ alkyl, C₂-C₁₀ alkene, C₁-C₆ alkoxy, C₃-C₈ cycloalkyl,C₂-C₈ heterocycloalkyl, C₄-C₁₀ aryl, C₄-C₁₀ aryloxy, C₁-C₈ heteroaryl,—(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₈ heteroaryl, —(CR′₅R′₆)_(m)—NR′₃R′₄,—(CR′₅R′₆)_(m)—C₃-C₈ heterocycloalkyl, —(CR′₅R′₆)_(m)—OR′₃,—(CR′₅R′₆)_(m)(O)COR′₃, —CO(O)R′₃, —CONR′₃R′₄, —NR′₃R′₄, —NR′₃(C(O)R′₄),—CH₂A where the compound of Chemical Formula 1 is “A”, wherein R′₃ andR′₄ are each independently hydrogen, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryl, —(CR′₅R′₆)_(m)—C₄-C₁₀ aryloxy,—(CR′₅R′₆)_(m)—C₁-C₁₀ heteroaryl, —CO(O)R″₃, or R′₃ and R′₄ takentogether form a cyclic structure of C₂-C₁₀ heterocycloalkyl, or a cyclicstructure of C₁-C₁₀ heteroaryl, the R′₅ and R′₆ are each independentlyhydrogen or C₁-C₃ alkyl, and the R″₃ is C₁-C₆ akyl.
 3. The methodaccording to claim 1, wherein the pharmaceutical composition comprises,as an active ingredient, a 1,2-naphthoquinone derivative compoundrepresented by the Chemical Formula 1 or a pharmaceutically acceptablesalt thereof, wherein the compound of the Chemical Formula 1 is one ofthe compounds represented below:


4. The method according to claim 3, wherein the pharmaceuticalcomposition comprises, as an active ingredient, a 1,2-naphthoquinonederivative compound represented by the Chemical Formula 1 or apharmaceutically acceptable salt thereof, wherein the compound iscompound 98 represented below:


5. The method according to claim 1, wherein the pharmaceuticalcomposition is one formulation selected from the group consisting ofpowders, granules, tablets, capsules, suspensions, emulsions, syrups,aerosols, epidermal formulations, suppositories, and sterile injectionsolutions.
 6. Compound 98 represented by the following structure, anoptical isomer thereof, or a pharmaceutically acceptable salt thereof: